FINAL

2007 Klamath River

Blue-Green Algae Summary

Report

Prepared by:

Ken Fetcho

Yurok Tribe

Environmental Program

July 2008

2

Acknowledgements

The Yurok Tribe Environmental Program (YTEP) would like to thank those that

contributed to the data collection efforts that occurred during the blue-green algae bloom

on the Klamath River during the 2007 monitoring season. AmeriCorps Watershed

Stewards Project Members Katie Cowan and Alex Wong were instrumental in collecting

samples and sending them to the appropriate labs. The Klamath Basin Tribal Water

Quality Workgroup and the State of CA were responsible for funding a large portion of

the analytical costs. YTEP would also like to thank Andy Lincoff at the USEPA Region

9 lab in Richmond, CA who processed microcystin samples at no cost. YTEP heartfully

thanks Jim Sweet at Aquatic Analysts in White Salmon, WA for sacrificing his personal

life to process our samples in a timely manner.

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Table of Contents

I. Introduction…………………………………………………….….………………..…4

II. Methods……………………………………………………………….…………..…..5

III. Site Selection……….………………..……………….………………………………6

IV. Quality Assurance….…………………………………………………………...….10

V. Results…………….………………………………………………………………….13

VI. Discussion……………………………………………...……………………………17

Figures

1 Map of monitoring locations ………………………………………………………….8

2 Map of Klamath River Estuary Monitoring Locations…………… ………………..9

3 Aphanizomenon flos-aquae levels, Klamath River Miles 44 to 0.5.. ………...….…16

4 Microcystis aeruginosa levels, Klamath River Miles 44 to 0.5 …………….………16

5 Microcystin levels, Klamath River Miles 44 to 0.5…………… ………….……..…17

Tables

1 Phytoplankton Replicate Results…………………….………….…………….…..…13

2 Microcystin Replicate Results …………………………………………….…….….…13

3 Phytoplankton Results Klamath and Trinity Rivers .……………….……....…..…14

4 Microcystis aeruginosa Results in edge water habitats in Klamath River Estuary.17

5 Microcystin Results for Klamath River …………………………………………….17

Literature Cited..…………………………………………………………………….…21

Appendix………………………………………………………………….………..……22

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I. Introduction

This report summarizes the occurrence and extent of the blue-green algae bloom on the

Klamath River within the Yurok Indian Reservation (YIR) boundaries in 2007. The

Karuk Tribe will be publishing a summary report that details the results for the algae and

nutrients conditions in Copco and Iron Gate Reservoirs. The Yurok Tribe Environmental

Program (YTEP), the Karuk Tribe, BOR and PacifiCorp collected water samples in 2007

indicating that the water quality in the Klamath River was negatively impacted by levels

of the cyanobacterium Microcystis aeruginosa (MSAE) and its resultant toxin,

microcystin; with some levels exceeding World Health Organization (WHO) risk

guidelines in all reaches of the Klamath River from downstream of Upper Klamath Lake

to the Klamath River Estuary. Water quality sampling performed in 2005 and 2006

detected MSAE and microcystin in the Klamath River below Iron Gate Dam. The Karuk

Tribe and YTEP were integral in collecting this data and sharing results with the water

quality regulators and the local community.

Information on Microcystis aeruginosa MSAE is a type of blue-green algae which, upon death and decomposition,

releases the hepatoxin microcystin, which can cause any of the following

reactions in humans and/or animals: rash, irritation, conjunctivitis, nausea,

vomiting, diarrhea, liver damage, tingling, numbness, paralysis, and death.

Microcystin bioaccumulates in the liver, organs, and to a certain degree the

muscle mass of living animals. Microcystin is not excreted by animals, and

dosage over time will eventually cause liver damage, decreased liver function and

increased liver size, and eventually death. Mortality in fish, domestic animals,

and humans has been recorded following exposure to microcystin resultant from

both single-dose events and long-term exposure.

The toxin microcystin can produce negative health effects from contact with

impaired waters, from incidental nasal/oral ingestion of impaired waters, and,

most seriously, from swallowing or drinking impaired waters. Microcystin can be

found in the organs and muscle meat of fish who feed in impaired waters.

Continued exposure to even low levels of microcystin can produce harmful

cumulative response in humans and animals. Because the timing for likely bloom

occurrence coincides with the annual salmon runs along the Klamath River, there

is significant cause for concern; Tribal subsistence fishers not only have increased

contact with impaired waters due to subsistence fishing methods, they are also

more frequently visiting impaired waters than recreational or sport users.

Initial bloom characteristics are green, green-blue, yellow, or whitish waters;

blooms of increasing severity can form algal mats or visible surface scum,

initially in nearshore areas and backwaters or where winds cause surface detritus

to accumulate, and steadily progressing to cover open areas of still or slow-

moving waters. Odor and taste changes may become noticeable at any time as

decomposition of organisms sets in, but should not be used as an indicator for

cellular mortality.

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Microcystin Toxin Information WHO has established minimum tolerance levels for recreational contact with

microcystin. Because of the time it takes to analyze water samples for the

presence of microcystin, WHO recommends the use of cell counts per milliliter of

water as a crude surrogate for concentrations of microcystin. However, because

the toxin is released as the organism decomposes, the risk from microcystin

presence in waters is at its greatest after the bloom has initially begun to

decompose and increases until well after the last cells are observed in samples.

WHO has set the following thresholds for MSAE/microcystin concentrations in

recreational waters:

Microcystis cells/milliliter Microcystin micrograms/liter

Low Risk: 20,000 4

Moderate Risk: 100,000 20

Severe Risk 10,000,000 or visible scum 200

The consumption limit for microcystin is set as 0.04 micrograms per kilogram of

bodyweight per day. However, because even the consumption of relatively low

doses of microcystin over time will damage the liver of animals, continued

consumption of known contaminated food sources is not recommended.

II. Methods

At each sample site, sample water was collected with a pre-rinsed churn splitter as

specified in the grab sample protocol located in Appendix B. The churn was rinsed three

times with distilled water followed by three rinses with site river water. Samples were

drawn in a moving portion of the river in an attempt to collect water samples to represent

the river as a whole. The churn splitter allowed for distribution of a homogenous water

mixture into sample bottles used for algal identification and enumeration and testing for

microcystin.

The sample bottle for determination of algal species contained Lugol's preservative and

the toxin sample was preserved by freezing the bottle. Both of these samples were drawn

from the same churn of water because they are complementary to one another. All

samples were labeled with the following information: date, time, sampler, sample site,

study name. The sample ID was comprised of a two or three digit site ID and the date

(e.g. TG090107).

If a sampling crew member identified an area along the river that had scum lines, an

additional sample was collected at this site. The sample was labeled appropriately and

photographs of the sample area were taken. Additional quality control measures were

included in the sampling. At one site per trip a replicate split sample was sent to the

laboratory to assess laboratory performance and to gain improved confidence in the data.

Environmental information was also recorded at the time water samples were collected.

The data included water temperature, pH, specific conductance, dissolved oxygen and

other observational notes. Water samples were also collected to be analyzed for the

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concentration of nutrient analytes and sent to Aquatic Research Inc. in Seattle,

Washington (WA). Chain-of-custody (COC) sheets were also filled out to document the

handling of the samples from the time of collection to the time of laboratory analysis.

This is a standard procedure for handling samples.

Water samples that were collected for algae speciation and enumeration were mailed

overnight to Aquatic Analysts in White Salmon, WA for analysis. Microscope slides are

prepared at the laboratory from each sample by filtering an appropriate aliquot of the

sample through a 0.45 micrometer membrane filter (APHA Standard Methods, 1992,

10200.D.2; McNabb, 1960). A section is cut out and placed on a glass slide with

immersion oil added to make the filter transparent, followed by placing a cover slip on

top, with nail polish applied to the periphery for permanency. Most algae are identified

by cross-referencing several taxonomic sources.

Algal units (defined as discrete particles - either cells, colonies, or filaments) are counted

along a measured transect of the microscope slide with a Zeiss standard microscope

(1000X, phase contrast). Algal units are measured accurately to 0.1 mm with a stage

micrometer. The algal densities are calculated from the area observed (transect length

times diameter of field of view), the effective filter area, and the volume of sample

filtered. Only those algae that were believed to be alive at the time of collection (intact

chloroplast) are counted. A minimum of 100 algal units are counted. (Standard Methods,

1992, 10200.F.2.c.). Average biovolume estimates of each species are obtained from

calculations of microscopic measurements of each alga. The number of cells per colony,

or the length of a filament, are recorded during sample analysis to arrive at biovolume per

unit-alga. Average biovolumes for algae are stored in a computer, and measurements are

verified for each sample analyzed.

Water samples that were collected for microcystin processing were stored in glass

containers and mailed on ice overnight to USEPA Region 9 lab in Richmond, California

(CA) for analysis using the enzyme linked immunosorbent assay (ELISA) method. These methods have been adapted to a commercial ELISA kit (Microcystin Plate Kit,

EP-022) that is produced by Envirologix, Inc. (Portland Maine), which USEPA Region

9 lab in Richmond, CA employs and measures total microcystin.

III. Site Selection

In general, the various sampling locations were chosen in order to represent the average

ambient water conditions throughout the water column. The sites listed below indicate

established sampling locations for the collection of water samples for nutrient analysis

and phytoplankton speciation and enumeration from May through October on a biweekly

interval.

Once the presence of MSAE was detected at sampling sites upstream of the YIR (July 10,

2007) additional samples were collected beginning on July 24, 2007 and continued

through October to test for the presence of microcystin. The Trinity River monitoring site

was sampled for nutrient analysis and algae speciation and enumeration on a monthly

basis in order to save resources. Biweekly sampling did not occur in the Trinity River

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because the routine algae speciation results showed that no toxicogenic cyanobacteria

were present. However, algae speciation and enumeration and toxin sampling event did

occur in the Trinity River on a monthly interval to maintain a long-term dataset and to

verify the hypothesis that toxicogenic cyanobacteria sources are from locations on the

Klamath River upstream of the YIR boundaries.

The Klamath River monitoring site below the confluence with the Trinity River (KBW)

was relocated downstream approximately four miles to the Klamath River above Tully

Creek monitoring site (TC) on September 5, 2007. This monitoring site was relocated

due to YTEP’s access being denied by the landowner. The data from both of these sites

are considered comparable and representative of the conditions downstream of the

Klamath and Trinity River confluence.

YTEP collected water samples for toxin and algae speciation analysis at the following

mainstem Klamath River locations (river miles are approximate):

• WE - Klamath River at Weitchpec (upstream of Trinity River) – RM 43.5

• KBW - Klamath River below Weitchpec RM 42.5

• TC – Klamath River Above Tully Creek – RM 38.5

• TG - Klamath River at Turwar Boat Ramp – RM 6

• LES - Lower Estuary Surface – RM 0.5

YTEP collected water samples for toxin and speciation analysis at the following major

tributary location:

• TR - Trinity River near mouth (above Klamath River confluence) – RM 0.5

YTEP did collect water samples in edge water habitats in the Klamath River Estuary at

various times during the 2007 monitoring season to determine if elevated levels of

toxicogenic cyanobacteria were present. These results are provided in this report

separately and are clearly identified as unique sampling locations (see figure 2).

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Figure 1. Map of phytoplankton and microcystin monitoring locations, 2007.

9

Figure 2. Map of phytoplankton monitoring locations including edge water

habitats in the Klamath River Estuary, 2007.

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IV. Quality Assurance

YTEP performs all surface water quality monitoring activities consistent with its Quality

Assurance Program Plan that was approved by the USEPA in April 2001. Quality

assurance/Quality control (QA/QC) of the collection, preparation and analysis of water

samples for microcystin and phytoplankton speciation and enumeration was achieved by

following a standard water sample collection protocol using a churn sampler and

submitting samples to labs that follow strict protocol that have QA/QC measures. All

field personnel that were involved in collection of water samples have been trained

appropriately by the Water Division Program Manager and are properly supervised to

ensure proper protocol is followed consistently throughout the monitoring season. Field

crews collecting samples ensured representativeness of samples by selecting free-flowing

water from established sampling locations and using a churn splitter to mix sample water

once collected. All samples were transported to the appropriate laboratories following

chain of custody procedures to ensure proper handling of the samples.

The collection and analysis of field replicate samples were performed on a monthly basis

to determine the labs’ precision of data. Field replicates were collected by splitting

samples in the field using the churn splitter. One of the split samples was sent with its’

associated split with a different ID code for analysis of both algae identification and

enumeration and microcystin so as to not alert lab staff of the fact that the samples were

replicates.

Data is thoroughly reviewed once received from the laboratory. YTEP is the primary

organization responsible for data review, although the professional laboratories analyzing

water quality samples will also note potential problems with outliers or other anomalies

in sample results. Information regarding QA/QC procedures for the laboratory is

available upon request. One hundred percent of laboratory-generated data was checked

on receipt by the Project Manager for consistency and acceptability, including whether

replicates are within specified targets and meet data quality objectives. Any unusual

values outside the range of norm will be flagged and all aspects of field data sheets,

shipping handling and laboratory handling and testing will be reviewed. Outliers will be

identified and removed from the dataset if deemed necessary by the QA Officer. Water

temperature, conductivity, pH and dissolved oxygen are measured in the field when

samples are collected and values of these hand-held measurements can be used to check

field conditions at the time of sampling.

The data manager will visually inspect all entered data sets to check for inconsistencies

with original field or laboratory data sheets. Where inconsistencies are encountered, data

will be re-entered and re-inspected until the entered data is found to be satisfactory or

results will be discarded. The Project Manager will maintain field datasheets and

notebooks in the event that the QA Officer needs to review any aspect of sampling for

QA/QC purposes.

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Phytoplankton

Not all of the phytoplankton replicate samples contained MSAE. Samples collected in

May, June and October did not contain MSAE, therefore, no relative percent differences

were calculated for these months. The common algae in each replicate did change rank

for most samples collected between May and October (see table 3). However, the same

species generally make up the bulk of the samples. Furthermore, the abundances

(whether density, biovolume, or Trophic State Index (TSI)) were similar for the

replicates. Considering the biological variability in this river the replicate results are not

significantly different. Discrepancy among replicates may also reflect how well the

churn was able to split particulates in samples.

The July QA replicate sample did contain low levels of MSAE (1,101 cells/ml) but the

primary sample did not report the presence of MSAE, therefore a RPD was not

calculated. The August QA replicate did contain MSAE, the primary sample reported

8,054 cells/ml and the replicate sample was 21,867 cells/ml, the RPD was 92.3%. The

September samples contained the highest levels of MSAE reported for the 2007 season.

The September primary sample was 90,764 cells/ml and the replicate sample was 34,133

cells/ml, the RPD was 90.7%.

The replicate results indicate that when MSAE is significantly present in water samples

the variability in replicate sample results increases. The variability of the results needs to

be viewed in light of the main objective of the phytoplankton sampling, which is to

inform the public if water contact poses a health risk. The August results have a high

RPD (92.3%) but both the primary and the replicate sample results are below the State of

CA recommended threshold to post a waterbody when MSAE exceeds 40,000 cells/ml.

The September results do pose a question in regards to deciding on whether to post a

waterbody or not.

The September results from additional sites located up-river from the site in which

replicate samples were collected (LES) in September aided YTEP in making the decision

to post the Klamath River within YIR boundaries following the guidance provided by the

State of California. In this case it was apparent that posting should occur because the

most upstream monitoring site on the YIR also exhibited high levels of MSAE. The

Klamath River site at Weitchpec that is above the Trinity River confluence had MSAE

cell densities of 80,016 cells/ml. YTEP was justified in posting due to the fact that two

sites on the reservation exhibited cell densities above the posting threshold and the fact

that samples collected upstream of the YIR by the Karuk Tribe reported levels at three

mainstem Klamath River monitoring sites ranging from 42,281 to 90,054 cells/ml

indicating that a significant risk was present to those who would have had contact with

the Klamath River downstream of Iron Gate Dam and could continue for some time.

Microcystin

QA replicate samples collected by both the Karuk Tribe and YTEP indicate that

the toxin results are valid and acceptable. All of the replicate samples submitted to the

lab generated RPD’s less than 25% (see table 2). These results are delivered after YTEP

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receives phytoplankton results, therefore, the toxin results were not used as the primary

source of information for determination of the need to post.

V. Results:

Phytoplankton QA Table 1. Phytoplankton results for the QA replicate samples collected by Karuk Tribe and YTEP, 2007.

Lab

Slide

ID

Station

ID Date

Total

Density

Total

Biovolume TSI Sp1 Sp1% Sp2 Sp2% Sp3 Sp3% Sp4 Sp4% Sp5 Sp5% #Spp

APF9

cells/ml

MSAE

cells/ml

KN03 TG 5/30/07 1,147 401,260 43.3 DTTN 21.7 NZFR 15.7 ACMN 13.0 RHCU 8.7 COPC 7.8 26 0 0

KN02 TG rep 5/30/07 1,028 324,550 41.7 DTTN 29.1 NZFR 14.6 ACMN 11.7 NZDS 5.8 NVCV 5.8 21 0 0

KP58P SC 6/26/07 3,750 477,457 44.5 ACMN 83.9 CMMN 5.6 CMAF 2.8 DTTN 1.4 NVCR 1.4 11 0 0

KP55P SC rep 6/26/07 3,769 478,245 44.5 ACMN 82 CMMN 7.4 CMSN 2.5 CMAF 1.6 NVCR 1.6 11 0 0

KN22 LES 7/24/07 706 70,594 31 KMXX 50.8 RDMN 22.6 SLMN 5.6 COPC 4.0 AKFL 3.2 15 0 0

KN24 LES rep 7/24/07 156 68,304 30.6 COPC 12.3 RDMN 12.3 EPSX 7.7 MSAE 7.7 SCQD 6.2 26 0 1,101

LA38P KRBI 8/21/07 5,450 7,040,486 63.9 APF9 93.6 RDMN 1.5 GFVT 1 COPC 1 MSAE 1 9 107,113 8,054

LA39P KRBI rep 8/21/07 5,631 5,532,622 62.2 APF9 82.5 NZPL 5.3 MSAE 3.9 CHXX 1.5 CXER 1.5 13 78,993 21,867

KN46 LES 9/18/07 1,738 1,074,461 50 RDMN 53.2 MSAE 12.6 CXER 4.5 COPC 4.5 NZFR 4.5 22 0 90,764

KN45 LES rep 9/18/07 1,321 537,601 45.4 RDMN 52.6 EPSX 7.8 MSAE 7.8 NZFR 5.2 SCQD 5.2 20 0 34,133

LF29P KRBI 10/16/07 237 219,195 38.9 RDMN 32.5 CXER 26.3 MLGR 10.5 COPC 9.6 GFAN 4.4 20 0 0

LF30P KRBI rep 10/16/07 505 206,473 38.5 RDMN 66.4 CXER 15.3 MLGR 3.6 COPC 2.9 GFAN 1.5 19 0 0 Key to Species Codes is located in Combined Species List located in Appendix A

Microcystin QA Table 2. Microcystin results for the QA replicate samples collected by Karuk Tribe and YTEP, 2007.

site ID primary result duplicate result difference RPD

LES072407 <1.8 <1.8 0.0 0.0

KRBI082107 5.4 4.7 0.7 13.9

LES091807 <1.8 <1.8 0.0 0.0

KRBI 101607 <0.18 <0.18 0.0 0.0

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Phytoplankton Table 3. Phytoplankton results for water samples collected in the Klamath River and mouth of Trinity River May - October 2007.

Site

ID Date

Total

Density

Total

Biovolume Sp1 Sp1% Sp2 Sp2% Sp3 Sp3% Sp4 Sp4% Sp5 Sp5% #Spp APF9

cells/ml

MSAE

cells/ml

ABX9

cells/ml

Oscillatoria

sp. cells/ml

WE 5/30/07 1,579 445,222 NZFR 20.5 ACMN 18.8 RHCU 11.6 DTTN 8.9 COPC 7.1 21 0 0 0 0

KBW 5/30/07 910 308,706 DTTN 16.8 ACMN 12.4 NZFR 9.7 RHCU 8.0 CMMN 6.2 29 0 0 0 0

TG 5/30/07 1,147 401,260 DTTN 21.7 NZFR 15.7 ACMN 13.0 RHCU 8.7 COPC 7.8 26 0 0 0 0

LES 5/30/07 1,039 281,086 DTTN 27.1 ACMN 17.8 NZFR 9.3 NZDS 5.4 STHN 5.4 27 0 0 0 0

TR 5/30/07 665 189,570 DTTN 58.8 ACMN 14.7 EPSX 4.9 NVCV 2.0 ACLC 2.0 20 0 0 0 0

WE 6/12/07 1,010 376,105 ACMN 20.4 COPC 16.5 RHCU 9.7 NZFR 8.7 CMAF 7.8 24 0 0 0 0

KBW 6/12/07 814 264,242 RDMN 15.8 NZFR 15.8 ACMN 12.3 COPC 8.8 DTTN 8.8 26 0 0 0 0

TG 6/12/07 2,372 1,037,347 DTTN 26.7 NZFR 17.8 ACMN 10.9 COPC 9.9 CMAF 8.9 16 0 0 0 0

LES 6/12/07 7,517 3,100,700 DTTN 24.0 NZFR 13.0 CMAF 12.0 RDMN 11.0 COPC 10.0 23 0 0 0 0

WE 6/26/07 631 223,815 COPC 30.9 ACMN 14.9 NZFR 9.6 RHCU 8.5 NVCV 4.3 22 0 0 0 0

KBW 6/26/07 844 404,883 COPC 29.0 ACMN 13.1 CMAF 8.4 RHCU 8.4 NZFR 6.5 27 0 0 0 0

TG 6/26/07 668 312,561 COPC 19.2 NZFR 12.1 DTTN 7.1 CMAF 7.1 ACMN 4.0 29 0 0 0 0

LES 6/26/07 374 111,803 COPC 23.3 NZFR 7.0 ACMN 7.0 CHXX 7.0 DTTN 7.0 24 0 0 0 0

TR 6/26/07 524 362,106 SNUL 20.0 COPC 19.0 DTTN 11.0 ACMN 8.0 GFAN 7.0 26 0 0 0 0

WE 7/10/07 395 205,770 COPC 42.1 SCQD 6.6 EPSX 6.6 SNUL 4.1 CMAF 4.1 22 0 0 0 0

KBW 7/10/07 413 266,618 COPC 30.9 CMAF 9.3 EPSX 7.2 SLMN 5.2 RHCU 5.2 30 0 0 0 0

TG 7/10/07 606 291,771 COPC 10.9 CMAF 9.8 ACMN 9.8 EPSX 8.7 SCQD 7.6 31 0 0 0 0

LES 7/10/07 262 133,065 COPC 15.1 ACMN 11.8 EPSX 9.7 CMAF 7.5 NZFR 6.5 25 0 0 0 0

WE 7/24/07 916 513,052 COPC 20.5 SCQD 15.4 EPSX 11.1 NZPL 5.1 SCAC 4.3 32 0 3,124 0 0

KBW 7/24/07 655 367,253 COPC 26.4 EPSX 16.0 SCQD 10.4 SNUL 5.7 NZFR 5.7 24 0 0 0 0

TG 7/24/07 929 509,862 SCQD 14.7 DTTN 9.8 RDMN 8.8 COPC 8.8 EPSX 5.9 32 0 0 0 0

LES 7/24/07 706 70,594 KMXX 50.8 RDMN 22.6 SLMN 5.6 COPC 4.0 AKFL 3.2 15 0 0 0 0

TR 7/24/07 264 133,177 COPC 34.2 EPSX 7.0 NZFR 6.1 CMAF 5.3 ACLC 4.4 29 0 0 46 0

WE 8/7/07 1,293 1,102,795 EPSX 43.4 COPC 10.1 NZFR 8.5 SNUL 6.2 APF9 6.2 20 1,473 6,013 0 0

KBW 8/7/07 1,062 751,149 EPSX 32.3 COPC 15.1 NVCR 6.5 NZFR 6.5 APF9 6.5 20 822 4,567 0 0

TG 8/7/07 741 771,305 EPSX 33.7 SNUL 8.9 COPC 6.9 NZFR 5.9 NZPC 5.9 27 0 0 0 0

LES 8/7/07 303 225,405 EPSX 15.7 APF9 12.4 RDMN 10.1 SNUL 6.7 AKFL 5.6 29 562 0 34 0 Key to Species Codes is located in Combined Species List located in Appendix A

APF9 = Aphanizomenon flos-aquae MSAE = Microcystis aeruginosa ABX9= Anabaena sp.

15

Table 3(contd.) Phytoplankton results for water samples collected in the Klamath River and mouth of Trinity River May - October 2007.

Site

ID Date

Total

Density

Total

Biovolume Sp1 Sp1% Sp2 Sp2% Sp3 Sp3% Sp4 Sp4% Sp5 Sp5% #Spp APF9

cells/ml

MSAE

cells/ml

ABX9

cells/ml

Oscillatoria

sp. cells/ml

WE 8/21/07 2,176 1,517,892 EPSX 20.0 NZFR 17.3 APF9 12.7 NVCR 6.4 SNUL 5.5 25 4,154 9,890 0 0

KBW 8/21/07 1,303 1,030,585 EPSX 32.7 NZFR 16.3 APF9 10.6 MSAE 7.7 COPC 7.7 18 2,067 18,040 0 0

TG 8/21/07 2,328 1,852,435 EPSX 27.0 NZFR 12.6 SNUL 8.1 SCQD 6.3 DTTN 5.4 25 629 4,195 210 0

LES 8/21/07 409 470,390 EPSX 25.0 DTTN 13.0 APF9 10.9 COPC 6.5 RDMN 6.5 26 712 4,003 0 0

TR 8/21/07 165 67,648 COPC 25.0 DTTN 13.1 SLMN 8.3 EPSX 8.3 NVCV 7.1 27 0 0 0 0

WE 9/5/07 1,811 1,099,073 EPSX 24.5 NZFR 18.9 COPC 7.5 RDMN 4.7 SNUL 3.8 25 0 7,517 0 0

TC 9/5/07 1,148 918,871 EPSX 27.3 COPC 9.1 SCQD 7.3 MSAE 6.4 NZFR 6.4 27 0 16,443 0 0

TG 9/5/07 2,875 2,459,548 EPSX 30.6 NZFR 10.5 SNUL 7.7 SCQD 5.7 NVCR 5.7 30 0 18,405 0 0

LES 9/5/07 847 404,360 RDMN 25.2 DTTN 11.3 CCMG 11.3 SCQD 10.4 EPSX 7.0 26 0 11,791 0 0

WE 9/18/07 1,358 1,294,346 COPC 22.3 EPSX 17.9 MSAE 13.4 NZFR 10.7 NVCR 6.3 25 0 80,016 0 0

TC 9/18/07 1,465 824,823 EPSX 20.0 COPC 19.1 NZFR 13.0 NVCV 7.8 MSAE 7.0 25 0 19,773 0 0

TG 9/18/07 2,405 1,003,685 NZFR 18.8 EPSX 17.0 RHCU 11.6 SCQD 6.3 MSAE 6.3 26 0 21,498 0 0

LES 9/18/07 1,738 1,074,461 RDMN 53.2 MSAE 12.6 CXER 4.5 COPC 4.5 NZFR 4.5 22 0 90,764 0 0

TR 9/18/07 98 35,960 DTTN 20.3 RDMN 10.9 EPSX 10.9 ACMN 7.8 COPC 7.8 24 0 0 0 0

WE 10/2/07 1,948 1,410,656 COPC 26.9 EPSX 20.4 NVCR 8.3 NVCV 6.5 NZFR 6.5 26 0 21,648 0 0

TC 10/2/07 1,337 920,406 COPC 27.0 EPSX 21.6 NZFR 9.0 NVCR 7.2 SNUL 5.4 22 0 7,228 0 0

TG 10/2/07 999 586,928 RDMN 26.7 COPC 15.2 EPSX 13.3 NZFR 9.5 DTTN 7.6 21 0 21,884 0 0

LES 10/2/07 395 259,235 COPC 14.5 DTTN 12.0 EPSX 10.8 NZFR 9.6 MSAE 9.6 27 0 13,777 0 0

WE 10/15/07 1,489 1,640,785 SNUL 22.8 COPC 14.9 NZFR 11.9 NVCR 9.9 DTVL 7.9 23 0 0 0 147

TC 10/15/07 838 679,823 SNUL 23.9 COPC 16.8 NVCR 10.6 DTTN 9.7 NZFR 7.1 21 0 0 0 0

TG 10/15/07 1,006 701,086 DTTN 25.3 COPC 17.2 EPSX 11.1 SNUL 10.1 DTVL 5.1 25 0 0 0 0

LES 10/15/07 272 196,572 DTTN 27.8 COPC 22.2 SNUL 12.2 EPSX 7.8 SNMZ 4.4 19 0 0 0 0

TR 10/15/07 261 117,991 DTTN 52.0 COPC 9.2 EPSX 8.2 SNUL 6.1 NZPC 5.1 19 0 0 0 0 Key to Species Codes is located in Combined Species List located in Appendix A

APF9 = Aphanizomenon flos-aquae MSAE = Microcystis aeruginosa ABX9= Anabaena sp.

Aphanizomenon flos-aquae Levels

in the Klamath River RM 44 to RM 0.5

2007

0

2,000

4,000

6,000

8,000

10,000

5/13/07

5/27/07

6/10/07

6/24/07

7/8/07

7/22/07

8/5/07

8/19/07

9/2/07

9/16/07

9/30/07

10/14/07

10/28/07

Date

ce

lls/m

l

WE RM 44

KBW RM 43

TC RM 39

TG RM 7

LES RM 0.5

Figure 3. Aphanizomenon flos-aquae levels for water samples collected in the Klamath River from

RM 44 to RM 0.5, May through October 2007.

Microcystis aeruginosa Levels

in the Klamath River RM190 to RM0.5

2007

0

10,000

20,000

30,000

40,000

50,000

60,000

70,000

80,000

90,000

100,000

5/13/2007

5/27/2007

6/10/2007

6/24/2007

7/8/2007

7/22/2007

8/5/2007

8/19/2007

9/2/2007

9/16/2007

9/30/2007

10/14/2007

10/28/2007

Date

cell

s/m

l

WE RM 44

KBW RM 43

TC RM 39

TG RM 7

LES RM 0.5

WHO Recreational

Low Risk

20,000 cells/ml

WHO Recreational

Moderate Risk

100,000 cells/ml

Posting Action Level

40,000 cells/ml

Figure 4. Microcystis aeruginosa levels for water samples collected in the Klamath River from RM 44 to

RM 0.5, May through October 2007.

17

Table 4. Microcystis aeruginosa levels for water samples collected in edgewater habitats located in the

Klamath River Estuary , May through September 2007.

Site ID Date APF9 cells/ml MSAE cells/ml ABX9 cells/ml

MES SC 5/21/2007 0 0 0

LES SS 7/30/2007 0 0 101

MES SC 9/12/2007 0 3,373,154* 410,000*

LES US Chute 9/12/2007 0 20,231 0

* Note: Microcystis counts calculated from observing an area 12 times greater than the rest of the sample.

APF9 = Aphanizomenon flos-aquae MSAE = Microcystis aeruginosa ABX9= Anabaena sp.

MES SC=Middle Estuary Surface Side Channel

LES SS=Lower Estuary Surface outlet of South Slough

LES US Chute=Lower Estuary Surface Upstream of Chute

Microcystin

Table 5. Microcystin results for water samples collected in the Klamath River from RM 44 to RM 0.5, July

to October 2007.

Total Microcystin Date

units: µg/L Site 7/24/2007 8/7/2007 8/21/2007 9/5/2007 9/18/2007 10/2/2007 10/15/2007USEPA Region 9 Lab ELISA

quanitiation limit: 1.8 µg/L WE <1.8 <1.8 1.8 <1.8 <1.8 <1.8 DNS

KBW SL <1.8 2.0 DNS DNS DNS DNS

TC DNS DNS DNS <1.8 <1.8 <1.8 DNS

TG <1.8 <1.8 <1.8 <1.8 <1.8 <1.8 DNS

LES <1.8 <1.8 <1.8 <1.8 <1.8 <1.8 DNS

TR <1.8 DNS <1.8 DNS <1.8 DNS DNS DNS = Did Not Sample

Microcystin Levels

in the Klamath River RM 44 to RM 0.5

2007

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

16.0

18.0

20.0

5/13/2007

5/27/2007

6/10/2007

6/24/2007

7/8/2007

7/22/2007

8/5/2007

8/19/2007

9/2/2007

9/16/2007

9/30/2007

10/14/2007

10/28/2007

Date

mic

rog

ram

s/L

WE RM 44

KBW RM 43

TC RM 39

LES RM 0.5

TG RM 7WHO Recreational

Low Risk 4 µg/L

WHO Recreational

Moderate Risk

20 µg/L

Posting Action

Level 8 µg/L

Figure 5. Microcystin levels for water samples collected in the Klamath River from RM 44 to RM 0.5, July

through October 2007.

18

VI. Discussion:

Phytoplankton

Aphanizomenon flos-aquae Aphanizomenon flos-aquae is a species of concern due to its ability to produce toxins and

its abundance in the reservoirs managed by PacifiCorp located upstream of the YIR.

Aphanizomenon flos-aquae was present in 7 of 50 samples collected in the Klamath River

reach located within the YIR from RM 44 to RM 0.5 (WE,KBW,TC,TG,LES) and was

not detected in any of the water samples collected at the Trinity River monitoring site.

Summary information of all algae species identified and enumerated in this river reach is

presented in Appendix A. Aphanizomenon flos-aquae ranked as the 17th

most dominant

species out of 105 total species when looking at the average percent density (1.2%).

Aphanizomenon flos-aquae was first detected on August 7, 2007 at the WE, KBW and

LES monitoring sites. Aphanizomenon flos-aquae continued to be present in the Klamath

River at multiple sampling sites on 8/21/07 and reached its highest level of 4,154 cells/ml

at the WE monitoring site. Aphanizomenon flos-aquae was not detected in the Klamath

River within the YIR during subsequent sampling events.

Microcystis aeruginosa MSAE was present in 19 of 50 samples collected in the Klamath River reach located

within the YIR from RM 44 to RM 0.5 (WE,KBW,TC,TG,LES) and was not detected in

any of the water samples collected at the Trinity River monitoring site. Summary

information of all algae species identified and enumerated in this river reach is presented

in Appendix A. MSAE ranked as the 13th

most dominant species when looking at the

average percent density (2.1%). MSAE was first detected on July 24th

, 2007 at the WE

sampling site. MSAE continued to be present in the Klamath River at multiple

monitoring sites through October 2, 2007. MSAE was not detected in the Klamath River

on the last monitoring event of the season that took place on October 15, 2007.

The highest density of MSAE occurred at the LES sampling site on September 18, 2007

and was measured at 90,764 cells/ml. This was the sampling event in which multiple

sites in the Klamath River within the YIR exceeded the cell density posting action level

of 40,000 cells/ml. These results triggered a response by YTEP to post notices informing

the public to avoid contact with the Klamath River. This posting took place in

conjunction with the State of CA and Humboldt County posting the river upstream of the

YIR.

These results indicate that MSAE was present in the Klamath River within the YIR for

over two months, with cell density and microcystin levels peaking near the middle of

September. The timing is of significance because of the presence of adult salmon and

steelhead migrating upstream during this time period. This is also a time of increased

cultural and recreational use of the Klamath River by both Tribal Members and sport

fishermen.

19

Estuary Edge Water Habitats

Supplemental water samples were collected in the Klamath River Estuary in May, July

and September to determine if toxicogencic cyanobacteria species were thriving in edge

water habitats (see figure 2 and table 5). Two samples were collected (May 21 and

September 12) in the middle estuary side channel that is located across from the Klamath

River Jet Boat Tour’s boat docks. One sample was collected at the outlet of the South

Slough in the lower estuary on July 30, 2007 after YTEP staff observed filamentous algae

growth and some surface accumulations that were in close proximity to macrophytes.

Another sample was collected in the lower estuary just upstream of the chute on

September 12, 2007 after the mouth had constricted and was causing the estuary water

level to increase.

These additional water samples have indicated that the edge water habitats contain

elevated levels of toxicogenic cyanobacteria when species in the free flowing river are

present. High levels of Anabaena spp. and MSAE, 410,000 and 3,373,154 cells/ml

respectively, were detected in the middle estuary when the routine river monitoring sites

were reporting elevated levels. It is important to note that for this particular sample

(MES SC 091207), it was important to have a reliable estimate of the Microcystis

density, but not the other algae present in the sample. The taxonomist analyzed a lot

more of the sample (12 times as much), but only counted Microcystis in that extra

analysis. The MSAE counts were back-calculated (divided by 12) to adjust for the extra

area observed on the microscope slide. These levels are the highest ever reported in the

Klamath River within the YIR. No toxin samples were collected during these additional

sampling events.

Anabaena spp. and Oscillatoria spp. Anabaena spp. and Oscillatoria spp. are also a species of concern due to their ability to

produce toxins. Anabaena spp. was present in 4 of 50 samples collected in the Klamath

River reach located within the YIR from RM 44 to RM 0.5 (WE,KBW,TC,TG,LES).

Summary information of all algae species identified and enumerated in this river reach is

presented in Appendix A. Anabaena spp was detected at low levels (highest level was

210 cells/ml at TG) at the routine monitoring sites during the scheduled events. The

highest recorded level of Anabaena spp. was 410,000 cells/ml which occurred on

September 12, 2007 when a sample was collected in edge water habitat located in a

middle estuary side channel.

It is interesting to note that one sample out of five collected at the Trinity River

monitoring site reported very low levels of Anabaena spp. at 46 cells/ml. This detection

potentially may have been a result of wildlife transporting cells from the Klamath River

due to the monitoring site location’s close proximity the Trinity River at the confluence.

This detection potentially may also have been a result of cross contamination at the lab or

by field personnel. YTEP’s blank QA samples in the past have showed very low

presence or no presence of algae cells. YTEP will continue to monitor phytoplankton

trends over time in the Trinity River and will respond appropriately to any increase in

toxicogenic cyanobacteria species.

20

Oscillatoria spp. was detected at the WE monitoring site on October 15, 2007 at low

levels. YTEP will continue to monitor phytoplankton trends over time in the Klamath

River and will respond appropriately to any increase in toxicogenic cyanobacteria

species.

Microcystin

Microcystin was present above the quantitation limit of 1.8 micrograms/Liter (µg/L) in 2

samples collected in the Klamath River reach located within the YIR from RM 44 to RM

0.5 (WE,KBW,TC,TG,LES). Microcystin was first detected in this river reach beginning

on August 21, 2007 at the WE and KBW sampling sites. Microcystin was not detected in

water samples that were collected in September or October. The highest reported

microcystin level in this river reach occurred at the WE sampling site on August 21, 2007

and was measured at 2.0 µg/L.

These results are interesting because microcystin levels are lower in 2007 when

compared to microcystin levels measured in 2006 even though samples collected in 2007

reported higher levels of MSAE. MSAE was also present later in the season in 2007

when compared to 2006 and different environmental variables were present, lower water

temperatures, less intense sunlight, shorter days of sunlight, etc. Past experimental

research has shown that that increases in surface temperature coupled with nutrient

loading could initiate a shift in dominance within the Microcystis population, causing

toxic cells to comprise a greater percentage of the total population (Davis and Gobler

2007). This type of information could add to the list of probable reasons why

microcystin measured with the ELISA kit were lower in 2007 when compared to 2006.

Another reason why the ELISA results were lower in 2007 was that other microcystin

congeners may have been present that the ELISA test kit were unable to detect. There

are over 50 known microcystin congeners and the ELISA test is designed to cross-react

with some congeners but not all that are known to exist.

21

Literature Cited

Davis, T. and Gobler C. 2007. The Effects of Temperature and Eutrophication on Toxic and Non-

Toxic Strains of Microcystis within New York Lakes. School of Marine and Atmospheric

Sciences, Stony Brook, NY, USA.

22

Appendix A

Table A-1. Combined Algae Species List for Klamath River Sites located at River Mile 44 to 0.5 (WE,

KBW, TC, TG, and LES) May to October, 2007.

50 samples total

# Algae Species Ave % Den # samples Code

1 Cocconeis placentula 15.4 50 COPC

2 Epithemia sorex 11.8 44 EPSX

3 Diatoma tenue 8.3 43 DTTN

4 Nitzschia frustulum 8.1 47 NZFR

5 Rhodomonas minuta 5.4 37 RDMN

6 Achnanthes minutissima 4.7 41 ACMN

7 Synedra ulna 4.1 39 SNUL

8 Rhoicosphenia curvata 3.1 42 RHCU

9 Scenedesmus quadricauda 2.9 36 SCQD

10 Navicula cryptocephala 2.7 44 NVCR

11 Cymbella affinis 2.4 28 CMAF

12 Navicula cryptocephala veneta 2.1 37 NVCV

13 Microcystis aeruginosa 2.1 19 MSAE

14 Cyclotella meneghiniana 1.6 27 CCMG

15 Cymbella sinuata 1.3 34 CMSN

16 Nitzschia paleacea 1.3 29 NZPC

17 Aphanizomenon flos-aquae 1.2 7 APF9

18 Diatoma vulgare 1.2 28 DTVL

19 Nitzschia palea 1.2 26 NZPL

20 Gomphonema angustatum 1.1 28 GFAN

21 Chromulina sp. 1.1 4 KMXX

22 Cymbella minuta 1.1 27 CMMN

23 Selenastrum minutum 1.1 19 SLMN

24 Ankistrodesmus falcatus 0.9 25 AKFL

25 Stephanodiscus hantzschii 0.8 14 STHN

26 Achnanthes lanceolata 0.8 21 ACLC

27 Nitzschia dissipata 0.8 20 NZDS

28 Fragilaria construens venter 0.6 14 FRCV

29 Gomphonema subclavatum 0.6 19 GFSB

30 Amphora perpusilla 0.6 21 AFPR

31 Chlamydomonas sp. 0.6 16 CHXX

32 Synedra mazamaensis 0.6 15 SNMZ

33 Gomphonema ventricosum 0.5 20 GFVT

34 Cryptomonas erosa 0.5 15 CXER

35 Nitzschia communis 0.5 17 NZCM

36 Navicula tripunctata 0.4 18 NVTP

37 Nitzschia acicularis 0.4 12 NZAC

38 Synedra tenera 0.4 4 SNTN

39 Nitzschia amphibia 0.4 14 NZAM

40 Fragilaria vaucheria 0.3 13 FRVA

41 Navicula decussis 0.3 13 NVDC

42 Nitzschia innominata 0.3 11 NZIN

43 Melosira granulata 0.3 10 MLGR

44 Gomphoneis herculeana 0.2 9 GSHR

45 Cyclotella pseudostelligera 0.2 3 CCPS

23

Table A-1(contd.). Combined Algae Species List for Klamath River Sites located at River Mile 44 to 0.5

(WE, KBW, TC, TG, and LES) May to October, 2007.

50 samples total

# Algae Species Ave % Den # samples Code

46 Gomphonema olivaceum 0.2 10 GFOM

47 Melosira varians 0.2 8 MLVR

48 Nitzschia sp. 0.2 8 NZXX

49 Scenedesmus acuminatus 0.2 6 SCAC

50 Sphaerocystis schroeteri 0.2 7 SFSR

51 Navicula gregaria 0.2 5 NVGR

52 Scenedesmus denticulatus 0.2 6 SCDT

53 Unidentified flagellate 0.1 3 MXFG

54 Cocconeis pediculus 0.1 3 COPD

55 Achnanthes linearis 0.1 5 ACLN

56 Nitzschia linearis 0.1 6 NZLN

57 Navicula sp. 0.1 5 NVXX

58 Navicula pupula 0.1 4 NVPP

59 Navicula graciloides 0.1 4 NVGC

60 Scenedesmus abundans 0.1 5 SCAB

61 Fragilaria crotonensis 0.1 5 FRCR

62 Fragilaria construens 0.1 4 FRCN

63 Cocconeis disculus 0.1 2 CODS

64 Gomphonema clevei 0.1 4 GFCL

65 Coelastrum microporum 0.1 3 CUMC

66 Anabaena sp. 0.1 4 ABX9

67 Tetraedron minimum 0.1 3 TEMN

68 Ulothrix sp. 0.1 3 ULXX

69 Gyrosigma spencerii 0.1 3 GYSP

70 Gomphonema tenellum 0.0 2 GFTN

71 Amphora ovalis 0.0 2 AFOV

72 Navicula viridula 0.0 2 NVVR

73 Synedra parasitica 0.0 2 SNPR

74 Cymbella mexicana 0.0 2 CMMX

75 Cladophora sp. 0.0 2 CFXX

76 Epithemia turgida 0.0 2 EPTR

77 Glenodinium sp. 0.0 1 GDXX

78 Pediastrum boryanum 0.0 2 PSBR

79 Navicula capitata 0.0 2 NVCP

80 Denticula elegans 0.0 2 DNEL

81 Fragilaria pinnata 0.0 2 FRPN

82 Cymbella microcephala 0.0 1 CMMC

83 Gloeocystis ampla 0.0 1 GLAM

84 Hantzschia amphioxys 0.0 1 HZAM

85 Pinnularia sp. 0.0 1 PLXX

86 Cymbella tumida 0.0 1 CMTM

87 Rhopalodia gibba 0.0 1 RPGB

88 Gomphonema sp. 0.0 1 GFXX

89 Fragilaria leptostauron 0.0 1 FRLP

90 Diatoma hiemale mesodon 0.0 1 DTHM

24

Table A-1(contd.). Combined Algae Species List for Klamath River Sites located at River Mile 44 to 0.5

(WE, KBW, TC, TG, and LES) May to October, 2007.

50 samples total

# Algae Species Ave % Den # samples Code

91 Nitzschia capitellata 0.0 1 NZCP

92 Kephyrion sp. 0.0 1 KFXX

93 Navicula radiosa 0.0 1 NVRD

94 Cyclotella ocellata 0.0 1 CCOC

95 Oscillatoria sp. 0.0 1 OSX9

96 Nitzschia microcephala 0.0 1 NZMC

97 Anomoeoneis vitrea 0.0 1 AOVT

98 Diploneis elliptica 0.0 1 DPEL

99 Melosira ambigua 0.0 1 MLAM

100 Rhopalodia musculus 0.0 1 RPMS

101 Synedra cyclopum 0.0 1 SNCY

102 Synedra socia 0.0 1 SNSC

103 Navicula menisculus upsaliensis 0.0 1 NVMU

104 Amphipleura pellucida 0.0 1 AMPL

105 Nitzschia volcanica 0.0 1 NZVL

25

Table A-2 Combined Algae Species List for Mouth of Major Tributary Site (TR) May to October, 2007.

5 samples total

# Algae Species Ave % Den # samples Code

1 Diatoma tenue 20.0 5 DTTN

2 Cocconeis placentula 19.0 5 COPC

3 Epithemia sorex 7.9 5 EPSX

4 Synedra ulna 6.1 5 SNUL

5 Achnanthes minutissima 4.8 5 ACMN

6 Navicula cryptocephala veneta 3.4 4 NVCV

7 Nitzschia frustulum 3.0 5 NZFR

8 Gomphonema angustatum 2.8 4 GFAN

9 Rhodomonas minuta 2.6 3 RDMN

10 Achnanthes lanceolata 2.2 3 ACLC

11 Nitzschia paleacea 2.1 5 NZPC

12 Cymbella sinuata 2.1 4 CMSN

13 Selenastrum minutum 1.8 2 SLMN

14 Rhoicosphenia curvata 1.8 4 RHCU

15 Cymbella affinis 1.5 3 CMAF

16 Nitzschia acicularis 1.4 3 NZAC

17 Cymbella minuta 1.3 3 CMMN

18 Navicula cryptocephala 1.1 3 NVCR

19 Nitzschia palea 1.1 4 NZPL

20 Navicula decussis 1.0 3 NVDC

21 Gomphonema subclavatum 1.0 2 GFSB

22 Nitzschia dissipata 1.0 2 NZDS

23 Ankistrodesmus falcatus 0.9 4 AKFL

24 Scenedesmus quadricauda 0.9 2 SCQD

25 Melosira varians 0.8 3 MLVR

26 Amphora perpusilla 0.7 3 AFPR

27 Gomphonema ventricosum 0.6 2 GFVT

28 Chlamydomonas sp. 0.4 2 CHXX

29 Gomphonema tenellum 0.4 2 GFTN

30 Diatoma vulgare 0.4 2 DTVL

31 Scenedesmus denticulatus 0.4 1 SCDT

32 Gomphonema olivaceum 0.4 2 GFOM

33 Achnanthes linearis 0.4 2 ACLN

34 Cocconeis pediculus 0.4 1 COPD

35 Nitzschia communis 0.3 1 NZCM

36 Cymbella microcephala 0.3 1 CMMC

37 Cryptomonas erosa 0.3 1 CXER

38 Nitzschia linearis 0.3 1 NZLN

39 Synedra mazamaensis 0.3 1 SNMZ

40 Gloeocystis ampla 0.3 1 GLAM

41 Tetraedron minimum 0.2 1 TEMN

42 Nitzschia sp. 0.2 1 NZXX

43 Navicula tripunctata 0.2 1 NVTP

44 Navicula pupula 0.2 1 NVPP

45 Ulothrix sp. 0.2 1 ULXX

46 Fragilaria leptostauron 0.2 1 FRLP

47 Pediastrum boryanum 0.2 1 PSBR

48 Nitzschia innominata 0.2 1 NZIN

49 Gomphonema clevei 0.2 1 GFCL

50 Amphipleura pellucida 0.2 1 AMPL

51 Anabaena sp. 0.2 1 ABX9

26

Appendix B

Grab Sample Protocol

‘Grab sampling’ refers to water samples obtained by dipping a collection

container into the upper layer of a body of water and collecting a water sample (USGS

File Report -00213). For quality assurance/quality control (QA/QC) purposes replicate,

and blank bottle sets will be prepared and collected for one site each sampling period.

These additional bottle sets will be handled, prepared and filled following the same

protocol used for regular bottle sets and samples. General water quality parameters will

also be measured with a freshly calibrated portable multi-probe water quality instrument

during grab samples and recorded onto data sheets.

Upon arrival at each site, the sampling churn will be rinsed three times with

deionized (D.I.) water. The goal of rinsing is ‘equipment decontamination – the removal

from equipment, residues from construction and machining and the removal of

substances adhering to equipment from previous exposure to environmental and other

media’ (USGS Open File Report 00213). After rinsing with D.I. water, the churn will be

rinsed three times with stream water. The churn is then fully submerged into the stream

and filled to the lid with sample water. Completely filling the churn allows for all

samples to be filled from one churn; thereby minimizing differences in water properties

and quality between samples.

Proper use of the churn guarantees the water is well mixed before the sample is

collected. The churn should be stirred at a uniform rate by raising or lowering the splitter

at approximately 9 inches per second (Bel-Art Products, 1993). This mixing must

continue while the bottles are being filled. If filling is stopped for some reason, the

stirring rate must be resumed before the next sample is drawn from the churn. As the

volume of water in the churn decreases, the round trip frequency increases as the velocity

of the churn splitter remains the same. Care must be taken to avoid breaking the surface

of the water as the splitter rises toward the top of the water in the churn.

Sample bottles and chemical preservatives used were provided by associated

laboratories and were considered sterile prior to field usage. Sample bottles without

chemical preservatives were rinsed with stream water from the churn 2-3 times before

filling with sample water. In the case of bottles that contained chemical preservatives,

bottles were not rinsed before sample collection and care was taken to avoid over-spillage

that would result in chemical preservative loss. Collected samples will be placed in

coolers on ice or dry ice for transport to contracted laboratories for analysis.

QA/QC – Replicate bottle set

To ensure laboratory and sampling accuracy, one site every sampling period was

randomly selected to receive one additional QA/QC bottle set. This bottle set contain

replicate water samples. Replicate samples are obtained using the same process as

regular samples. These are used to assure the laboratory maintains precision within

results.

All bottle sets are then placed on ice and are transported to the associated

laboratories. All grab samples were processed within 24 hours or within known

laboratory holding periods.

27

Bibliography

Bel-Art Products. Churn Sample Splitter Instructions, 37805 Series. Pequannock, NJ,

1993.

Eaton, Andrew D., Lenore S. Clesceri, and Arnold E. Greenberg., ed. Standard Methods

for the Examination of Water and Wastewater. 19th

Edition. Washington D.C.,

1995.

Lurry,D.L. and C.M. Kolbe. Interagency field manual for the collection of Water

Quality Data. USGS Publication, Open File Report 00-213.